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Journal articles on the topic 'RHBD'

Author: Grafiati

Published: 4 June 2021

Last updated: 5 February 2022

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1

Feng, Z., B. Jiang, J. Chandra, M. Ghannoum, S. Nelson, and A. Weinberg. "Human Beta-defensins: Differential Activity against Candidal Species and Regulation by Candida albicans." Journal of Dental Research 84, no. 5 (May 2005): 445–50. http://dx.doi.org/10.1177/154405910508400509.

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Oral epithelial cell-derived human beta-defensins-1, -2, and -3 participate in innate immune responses against Candida. We hypothesized that these peptides utilize several mechanisms for protection. Recombinant hBD-1 and -2 were produced with the use of an insect cell/baculovirus expression system, while rhBD-3 was expressed as a fusion protein in E. coli. RhBD-2 and -3 were more effective at killing the candidal species at low micromolar concentrations than was rhBD-1, except for C. glabrata. While this species was relatively resistant to rhBD fungicidal activity, its adherence to oral epithelial cells was strain-specifically inhibited by the rhBDs. C. albicans hyphae were important in regulating hBD2 and -3 mRNA expression in primary human oral epithelial cells. Confocal microscopy of rhBD-2-challenged C. albicans suggests disruption of the fungal membrane. Results support the hypothesis that hBDs control fungal colonization through hyphal induction, direct fungicidal activity, and inhibition of candidal adherence.

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2

Hamed, Ehab A., and Inhee Lee. "Categorization and SEU Fault Simulations of Radiation-Hardened-by-Design Flip-Flops." Electronics 10, no. 13 (June 30, 2021): 1572. http://dx.doi.org/10.3390/electronics10131572.

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In the previous three decades, many Radiation-Hardened-by-Design (RHBD) Flip-Flops (FFs) have been designed and improved to be immune to Single Event Upsets (SEUs). Their specifications are enhanced regarding soft error tolerance, area overhead, power consumption, and delay. In this review, previously presented RHBD FFs are classified into three categories with an overview of each category. Six well-known RHBD FFs architectures are simulated using a 180 nm CMOS process to show a fair comparison between them while the conventional Transmission Gate Flip-Flop (TGFF) is used as a reference design for this comparison. The results of the comparison are analyzed to give some important highlights about each design.

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3

Shuler, Robert L. "Porting and Scaling Strategies for Nanoscale CMOS RHBD." IEEE Transactions on Circuits and Systems I: Regular Papers 62, no. 12 (December 2015): 2856–63. http://dx.doi.org/10.1109/tcsi.2015.2495779.

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4

Cannon, Ethan H., Joe Tostenrude, Manuel Cabanas-Holmen, Barry Meaker, Charles Neathery, Mike Carson, and Roger Brees. "At-Speed SEE Testing of RHBD Embedded SRAMs." IEEE Transactions on Nuclear Science 60, no. 6 (December 2013): 4207–13. http://dx.doi.org/10.1109/tns.2013.2288307.

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5

Sarker, Md Arifur R., Seungwoo Jung, Adrian Ildefonso, Ani Khachatrian, Stephen P. Buchner, Dale McMorrow, Pauline Paki, John D. Cressler, and Ickhyun Song. "Mitigation of Single-Event Effects in SiGe-HBT Current-Mode Logic Circuits." Sensors 20, no. 9 (May 1, 2020): 2581. http://dx.doi.org/10.3390/s20092581.

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It has been known that negative feedback loops (internal and external) in a SiGe heterojunction bipolar transistors (HBT) DC current mirrors improve single-event transient (SET) response; both the peak transient current and the settling time significantly decrease. In the present work, we demonstrate how radiation hardening by design (RHBD) techniques utilized in DC bias blocks only (current mirrors) can also improve the SET response in AC signal paths of switching circuits (e.g., current-mode logic, CML) without any additional hardening in those AC signal paths. Four CML circuits both with and without RHBD current mirrors were fabricated in 130 nm SiGe HBT technology. Two existing RHBD techniques were employed separately in the current mirrors of the CML circuits: (1) applying internal negative feedback and (2) adding a large capacitor in a sensitive node. In addition, these methods are also combined to analyze the overall SET performance. The single-event transients of the fabricated circuits were captured under the two-photon-absorption laser-induced single-event environment. The measurement data clearly show significant improvements in SET response in the AC signal paths of the CML circuits by using the two radiation hardening techniques applied only in DC current mirrors. The peak output transient current is notably reduced, and the settling time upon a laser strike is shortened significantly.

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6

Blaine, R. W., S. E. Armstrong, J. S. Kauppila, N. M. Atkinson, B. D. Olson, W. T. Holman, and L. W. Massengill. "RHBD Bias Circuits Utilizing Sensitive Node Active Charge Cancellation." IEEE Transactions on Nuclear Science 58, no. 6 (December 2011): 3060–66. http://dx.doi.org/10.1109/tns.2011.2171365.

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7

Bals, Robert, Christiane Lang, Daniel J. Weiner, Claus Vogelmeier, Ulrich Welsch, and James M. Wilson. "Rhesus Monkey (Macaca mulatta) Mucosal Antimicrobial Peptides Are Close Homologues of Human Molecules." Clinical Diagnostic Laboratory Immunology 8, no. 2 (March 1, 2001): 370–75. http://dx.doi.org/10.1128/cdli.8.2.370-375.2001.

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ABSTRACT One component of host defense at mucosal surfaces appears to be epithelium-derived antimicrobial peptides. Molecules of the defensin and cathelicidin families have been studied in several species, including human and mouse. We describe in this report the identification and characterization of rhesus monkey homologues of human mucosal antimicrobial peptides. Using reverse transcriptase PCR methodology, we cloned the cDNAs of rhesus monkey β-defensin 1 and 2 (rhBD-1 and rhBD-2) and rhesus monkey LL-37/CAP-18 (rhLL-37/rhCAP-18). The predicted amino acid sequences showed a high degree of homology to the human molecules. The expression of the monkey antimicrobial peptides was analyzed using immunohistochemistry with three polyclonal antibodies to the human molecules. As in humans, rhesus monkey antimicrobial peptides are expressed in epithelia of various organs. The present study demonstrates that β-defensins and cathelicidins of rhesus monkeys are close homologues to the human molecules and indicate that nonhuman primates represent valid model organisms to study innate immune functions.

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8

Ren, Yi, Li Chen, and Jinshun Bi. "An RHBD Bandgap Reference Utilizing Single Event Transient Isolation Technique." IEEE Transactions on Nuclear Science 63, no. 3 (June 2016): 1927–33. http://dx.doi.org/10.1109/tns.2016.2554104.

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9

Han, Zhiwei, Liang Wang, Suge Yue, Bing Han, and Shougang Du. "Analysis and RHBD technique of single event transients in PLLs." Journal of Semiconductors 36, no. 11 (November 2015): 115001. http://dx.doi.org/10.1088/1674-4926/36/11/115001.

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10

Díez-Acereda, V., Sunil L. Khemchandani, J. del Pino, and S. Mateos-Angulo. "RHBD Techniques to Mitigate SEU and SET in CMOS Frequency Synthesizers." Electronics 8, no. 6 (June 19, 2019): 690. http://dx.doi.org/10.3390/electronics8060690.

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This paper presents a thorough study of radiation effects on a frequency synthesizer designed in a 0.18 μ m CMOS technology. In CMOS devices, the effect of a high energy particle impact can be modeled by a current pulse connected to the drain of the transistors. The effects of SET (single event transient) and SEU (single event upset) were analyzed connecting current pulses to the drains of all the transistors and analyzing the amplitude variations and phase shifts obtained at the output nodes. Following this procedure, the most sensitive circuits were detected. This paper proposes a combination of radiation hardening-by-design techniques (RHBD) such as resistor–capacitor (RC) filtering or local circuit-redundancy to mitigate the effects of radiation. The proposed modifications make the frequency synthesizer more robust against radiation.

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11

Atkinson, Blaine, Kauppila, Armstrong, T. Daniel Loveless, Hooten, Holman, Massengill, and Warner. "RHBD Technique for Single-Event Charge Cancellation in Folded-Cascode Amplifiers." IEEE Transactions on Nuclear Science 60, no. 4 (August 2013): 2756–61. http://dx.doi.org/10.1109/tns.2013.2240316.

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12

Rodbell, Kenneth P., David F. Heidel, Jonathan A. Pellish, Paul W. Marshall, Henry H. K. Tang, Conal E. Murray, Kenneth A. LaBel, et al. "32 and 45 nm Radiation-Hardened-by-Design (RHBD) SOI Latches." IEEE Transactions on Nuclear Science 58, no. 6 (December 2011): 2702–10. http://dx.doi.org/10.1109/tns.2011.2171715.

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13

Aguiar, Y. Q., F. Wrobel, J. L. Autran, F. L. Kastensmidt, P. Leroux, F. Saigne, V. Pouget, and A. D. Touboul. "Exploiting Transistor Folding Layout as RHBD Technique Against Single-Event Transients." IEEE Transactions on Nuclear Science 67, no. 7 (July 2020): 1581–89. http://dx.doi.org/10.1109/tns.2020.3003166.

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14

HOLMAN, W. T. "RADIATION-TOLERANT DESIGN FOR HIGH PERFORMANCE MIXED-SIGNAL CIRCUITS." International Journal of High Speed Electronics and Systems 14, no. 02 (June 2004): 353–66. http://dx.doi.org/10.1142/s0129156404002405.

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Modern semiconductor processes can provide significant intrinsic hardness against radiation effects in digital and analog circuits. Current design techniques using commercial processes for radiation-tolerant integrated circuits are summarized, with an emphasis on their application in high performance mixed-signal circuits and systems. Examples of "radiation hardened by design" (RHBD) methodologies are illustrated for reducing the vulnerability of circuits and components to total dose, single-event, and dose-rate effects.

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15

Balasubramanian, A., B. L. Bhuva, J. D. Black, and L. W. Massengill. "RHBD techniques for mitigating effects of single-event hits using guard-gates." IEEE Transactions on Nuclear Science 52, no. 6 (December 2005): 2531–35. http://dx.doi.org/10.1109/tns.2005.860719.

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16

Dohar, Suraj Singh, Siddharth R. K., Vasantha M. H., and Nithin Kumar Y. B. "A 1.2 V, Highly Reliable RHBD 10T SRAM Cell for Aerospace Application." IEEE Transactions on Electron Devices 68, no. 5 (May 2021): 2265–70. http://dx.doi.org/10.1109/ted.2021.3064899.

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17

Sutton, Akil K., Marco Bellini, John D. Cressler, Jonathon A. Pellish, Robert A. Reed, Paul W. Marshall, Guofu Niu, Gyorgy Vizkelethy, Marek Turowski, and Ashok Raman. "An Evaluation of Transistor-Layout RHBD Techniques for SEE Mitigation in SiGe HBTs." IEEE Transactions on Nuclear Science 54, no. 6 (December 2007): 2044–52. http://dx.doi.org/10.1109/tns.2007.908697.

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18

Cabanas-Holmen, M., E. H. Cannon, A. Kleinosowski, J. Ballast, J. Killens, and J. Socha. "Clock and Reset Transients in a 90 nm RHBD Single-Core Tilera Processor." IEEE Transactions on Nuclear Science 56, no. 6 (December 2009): 3505–10. http://dx.doi.org/10.1109/tns.2009.2034314.

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19

Yeargin, Susan, Amy L. McKenzie, Lindsey E. Eberman, J. Derek Kingsley, David J. Dziedzicki, and Patrick Yoder. "Physiological and Perceived Effects of Forearm or Head Cooling During Simulated Firefighting Activity and Rehabilitation." Journal of Athletic Training 51, no. 11 (November 1, 2016): 927–35. http://dx.doi.org/10.4085/1062-6050-51.10.09.

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Context: Cooling devices aim to protect firefighters by attenuating a rise in body temperature. Devices for head cooling (HC) while firefighting and forearm cooling (FC) during rehabilitation (RHB) intervals are commonly marketed, but research regarding their efficacy is limited. Objective: To investigate the physiological and perceived effects of HC and FC during firefighting drills and RHB. Design: Randomized controlled clinical trial. Setting: Firefighter training center. Patients or Other Participants: Twenty-seven male career firefighters (age = 39 ± 7 years; height = 169 ± 7 cm; weight = 95.4 ± 16.8 kg). Intervention(s): Firefighters were randomly assigned to 1 condition: HC (n = 9), in which participants completed drills wearing a cold gel pack inside their helmet; FC (n = 8), in which participants sat on a collapsible chair with water-immersion arm troughs during RHB; or control (n = 10), in which participants used no cooling devices. Firefighters completed four 15-minute drills (D1−D4) wearing full bunker gear and breathing apparatus. Participants had a 15-min RHB after D2 (RHB1) and D4 (RHB2). Main Outcome Measure(s): Change (Δ) in gastrointestinal temperature (TGI), heart rate (HR), physiological strain index, and perceived thermal sensation. Results: The TGI increased similarly in the HC and control groups, respectively (D1: 0.57°C ± 0.41°C, 0.73°C ± 0.30°C; D2: 0.92°C ± 0.28°C, 0.85°C ± 0.27°C; D3: −0.37°C ± 0.34°C, −0.01°C ± 0.72°C; D4: 0.25°C ± 0.42°C, 0.57°C ± 0.26°C; P > .05). The ΔHR, Δ physiological strain index, and Δ thermal sensation were similar between the HC and control groups during drills (P > .05). The FC group demonstrated a decreased TGI compared with the control group after RHB1 (−1.61°C ± 0.35°C versus −0.23°C ± 0.34°C; P < .001) and RHB2 (−1.40°C ± 0.38°C versus −0.38°C ± 0.24°C; P < .001). The physiological strain index score decreased in the FC group compared with the control group after RHB1 (−7.9 ± 1.3 versus −2.6 ± 1.7; P < .001) and RHB2 (−7.9 ± 1.6 versus −3.6 ± 1.1; P < .001), but no differences between groups were demonstrated for ΔHR or Δ thermal sensation (P > .05). Conclusions: The HC did not attenuate rises in physiological or perceptual variables during firefighting drills. The FC effectively reduced TGI and the physiological strain index score but not HR or thermal sensation during RHB. Clinicians and firefighters should not recommend the use of HC during firefighting but can consider using FC during RHB intervals in the field.

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20

Zhao, Yuanfu, Liang Wang, Suge Yue, Dan Wang, Xinyuan Zhao, Yongshu Sun, Dongqiang Li, et al. "SEU and SET of 65 Bulk CMOS Flip-flops and Their Implications for RHBD." IEEE Transactions on Nuclear Science 62, no. 6 (December 2015): 2666–72. http://dx.doi.org/10.1109/tns.2015.2490552.

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21

Krithivasan, Ramkumar, Paul W. Marshall, Mustayeen Nayeem, Akil K. Sutton, Wei-Min Kuo, Becca M. Haugerud, Laleh Najafizadeh, et al. "Application of RHBD Techniques to SEU Hardening of Third-Generation SiGe HBT Logic Circuits." IEEE Transactions on Nuclear Science 53, no. 6 (December 2006): 3400–3407. http://dx.doi.org/10.1109/tns.2006.885379.

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22

Hindman, Nathan D., Ziyan Wang, Lawrence T. Clark, and David R. Allee. "Experimentally Measured Input Referred Voltage Offsets and Kickback Noise in RHBD Analog Comparator Arrays." IEEE Transactions on Nuclear Science 54, no. 6 (December 2007): 2073–79. http://dx.doi.org/10.1109/tns.2007.908654.

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23

Mohr, Karl C., Lawrence T. Clark, and Keith E. Holbert. "A 130-nm RHBD SRAM With High Speed SET and Area Efficient TID Mitigation." IEEE Transactions on Nuclear Science 54, no. 6 (December 2007): 2092–99. http://dx.doi.org/10.1109/tns.2007.910867.

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24

Battezzati, N., F. Margaglia, M. Violante, F. Decuzzi, D. Merodio Codinachs, and B. Bancelin. "Application-Oriented SEU Cross-Section of a Processor Soft Core for Atmel RHBD FPGAs." IEEE Transactions on Nuclear Science 58, no. 3 (June 2011): 987–92. http://dx.doi.org/10.1109/tns.2010.2103326.

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25

Guo, Jing, Lei Zhu, Yu Sun, Huiliang Cao, Hai Huang, Tianqi Wang, Chunhua Qi, et al. "Design of Area-Efficient and Highly Reliable RHBD 10T Memory Cell for Aerospace Applications." IEEE Transactions on Very Large Scale Integration (VLSI) Systems 26, no. 5 (May 2018): 991–94. http://dx.doi.org/10.1109/tvlsi.2017.2788439.

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26

Quilligan, G., and S. Aslam. "TID and Heavy-Ion Performance of an RHBD Multichannel Digitizer in 180-nm CMOS." IEEE Transactions on Nuclear Science 68, no. 7 (July 2021): 1414–22. http://dx.doi.org/10.1109/tns.2021.3080179.

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27

Liu, Jingtian, Qian Sun, Bin Liang, Jianjun Chen, Yaqing Chi, and Yang Guo. "Bulk Bias as an Analog Single-Event Transient Mitigation Technique with Negligible Penalty." Electronics 9, no. 1 (December 25, 2019): 27. http://dx.doi.org/10.3390/electronics9010027.

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In analog circuit design, the bulks of MOSFETs can be tied to their respective sources to remove body effect. This paper models and analyzes the sensitivity of single-event transients (SETs) in common source (CS) amplifier with bulk tied to source (BTS) in 40 nm twin-well bulk CMOS technology. The simulation results present that the proposed BTS radiation-hardened-by-design (RHBD) technique can reduce charge collection and suppress the SET induced perturbation effectively in various input conditions of the circuit. The detailed analysis shows that the mitigation of SET is primarily due to the forward-bias of bulk potential. This technique is universally applicable in radiation-hardening design for analog circuits with negligible penalty.

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28

Blaine, R. W., N. M. Atkinson, J. S. Kauppila, S. E. Armstrong, N. C. Hooten, J. H. Warner, W. T. Holman, and L. W. Massengill. "Differential Charge Cancellation (DCC) Layout as an RHBD Technique for Bulk CMOS Differential Circuit Design." IEEE Transactions on Nuclear Science 59, no. 6 (December 2012): 2867–71. http://dx.doi.org/10.1109/tns.2012.2222441.

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29

Jun, Bongim, Akil K. Sutton, Ryan M. Diestelhorst, Gregory J. Duperon, John D. Cressler, Jeffrey D. Black, Tim Haeffner, et al. "The Application of RHBD to n-MOSFETs Intended for Use in Cryogenic-Temperature Radiation Environments." IEEE Transactions on Nuclear Science 54, no. 6 (December 2007): 2100–2105. http://dx.doi.org/10.1109/tns.2007.910123.

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30

Guo, Jing, Lei Zhu, Wenyi Liu, Hai Huang, Shanshan Liu, Tianqi Wang, Liyi Xiao, and Zhigang Mao. "Novel Radiation-Hardened-by-Design (RHBD) 12T Memory Cell for Aerospace Applications in Nanoscale CMOS Technology." IEEE Transactions on Very Large Scale Integration (VLSI) Systems 25, no. 5 (May 2017): 1593–600. http://dx.doi.org/10.1109/tvlsi.2016.2645282.

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31

Bogorad, Alexander L., Justin J. Likar, Robert E. Lombardi, Stephen E. Stone, and Roman Herschitz. "On-Orbit Error Rates of RHBD SRAMs: Comparison of Calculation Techniques and Space Environmental Models With Observed Performance." IEEE Transactions on Nuclear Science 58, no. 6 (December 2011): 2804–6. http://dx.doi.org/10.1109/tns.2011.2167242.

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32

Andreou, Charalambos M., Diego Miguel González-Castaño, Simone Gerardin, Marta Bagatin, Faustino Gómez Rodriguez, Alessandro Paccagnella, Alexander V. Prokofiev, et al. "Low-Power, Subthreshold Reference Circuits for the Space Environment: Evaluated with γ-rays, X-rays, Protons and Heavy Ions." Electronics 8, no. 5 (May 21, 2019): 562. http://dx.doi.org/10.3390/electronics8050562.

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The radiation tolerance of subthreshold reference circuits for space microelectronics is presented. The assessment is supported by measured results of total ionization dose and single event transient radiation-induced effects under γ -rays, X-rays, protons and heavy ions (silicon, krypton and xenon). A high total irradiation dose with different radiation sources was used to evaluate the proposed topologies for a wide range of applications operating in harsh environments similar to the space environment. The proposed custom designed integrated circuits (IC) circuits utilize only CMOS transistors, operating in the subthreshold regime, and poly-silicon resistors without using any external components such as compensation capacitors. The circuits are radiation hardened by design (RHBD) and they were fabricated using TowerJazz Semiconductor’s 0.18 μm standard CMOS technology. The proposed voltage references are shown to be suitable for high-precision and low-power space applications. It is demonstrated that radiation hardened microelectronics operating in subthreshold regime are promising candidates for significantly reducing the size and cost of space missions due to reduced energy requirements.

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33

Monda, Danilo, Gabriele Ciarpi, and Sergio Saponara. "Diagnosis of Faults Induced by Radiation and Circuit-Level Design Mitigation Techniques: Experience from VCO and High-Speed Driver CMOS ICs Case Studies." Electronics 10, no. 17 (September 3, 2021): 2144. http://dx.doi.org/10.3390/electronics10172144.

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In this paper, we discuss the diagnosis of particle-induced failures in harsh environments such as space and high-energy physics. To address these effects, simulation-before-test and simulation-after-test can be the key points in choosing which radiation hardening by design (RHBD) techniques can be implemented to mitigate or prevent failures. Despite the fact that total ionising dose (TID) has slow but destructive effects overtime on silicon devices, single-event effect (SEE) impulsively disrupts the typical operation of a circuit with temporary or permanent effects. The recently released SpaceFibre protocol drives the current requirements for space applications, and the future upgrade of the LHC experiment scheduled by CERN will require a redesign of the electronic front-end to sustain a radiation level up to the 1 Grad TID level. The effects that these two environments have on two different architectures for high-radiation and high-frequency data transmission are reported, and the efficiency of the mitigation techniques implemented, based on simulations and measurement tests, in the commercial 65 nm technology, are exploited.

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34

Adachi, K., CH Lai, P. Konitzer, M. Donahee, A. Campbell, and S. Surrey. "Crystallization of recombinant hemoglobins with basic amino acid substitutions (Lys and Arg) at the beta 6 position." Blood 84, no. 4 (August 15, 1994): 1309–13. http://dx.doi.org/10.1182/blood.v84.4.1309.1309.

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Abstract We have produced recombinant hemoglobins (rHbs) alpha 2 beta 2(6Glu-- >Lys) (rHb beta E6K) and alpha 2 beta 2(6Glu-->Arg) (rHb beta E6R) using a yeast expression system coupled with a polymerase chain reaction (PCR)-based mutagenesis strategy for studies focused on defining determinants that facilitate crystallization of Hb C (alpha 2 beta 2(6Lys)). rHb beta E6K had the same electrophoretic mobility as native human Hb C, whereas rHb beta E6R migrated slightly slower than Hb C on cellulose acetate electrophoresis. The carbonmonoxy (CO) forms of rHb beta E6K and rHb beta E6R formed tetrahedral crystals in vitro in 2.3 mol/L phosphate buffer just like native Hb C. The Hb concentration required for crystallization of CO-rHb beta E6R was lower than that of CO-rHb beta E6K, suggesting that stronger basic amino acids at the beta 6 position accelerate crystallization of Hb. However, the size of rHb beta E6R crystals was smaller than that of rHb beta E6K. Crystallization of native Hb C and both rHbs was inhibited by Hb F. These results suggest that alpha 2 beta gamma-heterohybrids that have basic amino acids at the beta 6 position behave similarly and are unable to crystallize like Hb C.

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35

Adachi, K., CH Lai, P. Konitzer, M. Donahee, A. Campbell, and S. Surrey. "Crystallization of recombinant hemoglobins with basic amino acid substitutions (Lys and Arg) at the beta 6 position." Blood 84, no. 4 (August 15, 1994): 1309–13. http://dx.doi.org/10.1182/blood.v84.4.1309.bloodjournal8441309.

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We have produced recombinant hemoglobins (rHbs) alpha 2 beta 2(6Glu-- >Lys) (rHb beta E6K) and alpha 2 beta 2(6Glu-->Arg) (rHb beta E6R) using a yeast expression system coupled with a polymerase chain reaction (PCR)-based mutagenesis strategy for studies focused on defining determinants that facilitate crystallization of Hb C (alpha 2 beta 2(6Lys)). rHb beta E6K had the same electrophoretic mobility as native human Hb C, whereas rHb beta E6R migrated slightly slower than Hb C on cellulose acetate electrophoresis. The carbonmonoxy (CO) forms of rHb beta E6K and rHb beta E6R formed tetrahedral crystals in vitro in 2.3 mol/L phosphate buffer just like native Hb C. The Hb concentration required for crystallization of CO-rHb beta E6R was lower than that of CO-rHb beta E6K, suggesting that stronger basic amino acids at the beta 6 position accelerate crystallization of Hb. However, the size of rHb beta E6R crystals was smaller than that of rHb beta E6K. Crystallization of native Hb C and both rHbs was inhibited by Hb F. These results suggest that alpha 2 beta gamma-heterohybrids that have basic amino acids at the beta 6 position behave similarly and are unable to crystallize like Hb C.

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36

Banteywalu, Solomon, Baseem Khan, Valentijn De Smedt, and Paul Leroux. "A Novel Modular Radiation Hardening Approach Applied to a Synchronous Buck Converter." Electronics 8, no. 5 (May 8, 2019): 513. http://dx.doi.org/10.3390/electronics8050513.

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Radiation and extreme temperature are the main inhibitors for the use of electronic devices in space applications. Radiation challenges the normal and stable operation of DC-DC converters, used as power supply for onboard systems in satellites and spacecrafts. In this situation, special design techniques known as radiation hardening or radiation tolerant designs have to be employed. In this work, a module level design approach for radiation hardening is addressed. A module in this sense is a constituent of a digital controller, which includes an analog to digital converter (ADC), a digital proportional-integral-derivative (PID) controller, and a digital pulse width modulator (DPWM). As a new Radiation Hardening by Design technique (RHBD), a four module redundancy technique is proposed and applied to the digital voltage mode controller driving a synchronous buck converter, which has been implemented as hardware-in-the-loop (HIL) simulation block in MATLAB/Simulink using Xilinx system generator based on the Zynq-7000 development board (ZYBO). The technique is compared, for reliability and hardware resources requirement, with triple modular redundancy (TMR), five modular redundancy (FMR) and the modified triplex–duplex architecture. Furthermore, radiation induced failures are emulated by switching all duplicated modules inputs to different signals, or to ground during simulation. The simulation results show that the proposed technique has 25% and 30%longer expected life compared to TMR and FMR techniques, respectively, and has the lowest hardware resource requirement compared to FMR and the modified triplex–duplex techniques.

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37

Weickert, Michael J., Maria Pagratis, Christopher B. Glascock, and Richard Blackmore. "A Mutation That Improves Soluble Recombinant Hemoglobin Accumulation in Escherichia coli in Heme Excess." Applied and Environmental Microbiology 65, no. 2 (February 1, 1999): 640–47. http://dx.doi.org/10.1128/aem.65.2.640-647.1999.

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ABSTRACT High-level expression of soluble recombinant human hemoglobin (rHb) in Escherichia coli was obtained with several hemoglobin variants. Under identical conditions, two rHbs containing the Presbyterian mutation (Asn-108→Lys) in β-globin accumulated to approximately twofold less soluble globin than rHbs containing the corresponding wild-type β-globin subunit accumulated. The β-globin Providence(asp) mutation (Lys-82→Asp) significantly improved soluble rHb accumulation compared to the wild-type β-globin subunit and restored soluble accumulation of rHbs containing the Presbyterian mutation to wild-type levels. The Providenceasp substitution introduced a negatively charged residue into the normally cationic 2,3-bisphosphoglycerate binding pocket, potentially reducing the electrostatic repulsion in the absence of the polyanion. The average soluble globin accumulation when there was coexpression of di-α-globin and β-Lys-82→Asp-globin (rHb9.1) and heme was present in at least a threefold molar excess was 36% ± 3% of the soluble cell protein in E. coli. The average total accumulation (soluble globin plus insoluble globin) was 56% ± 7% of the soluble cell protein. Fermentations yielded 6.0 ± 0.3 g of soluble rHb9.1 per liter 16 h after induction and 6.4 ± 0.2 g/liter 24 h after induction. The average total globin yield was 9.4 g/liter 16 h after induction. High-level accumulation of soluble rHb in E. coli depends on culture conditions, the protein sequence, and the molar ratio of the heme cofactor added.

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Panepinto, John C., Brian G. Oliver, Jarrod R. Fortwendel, Darcey L. H. Smith, David S. Askew, and Judith C. Rhodes. "Deletion of the Aspergillus fumigatus Gene Encoding the Ras-Related Protein RhbA Reduces Virulence in a Model of Invasive Pulmonary Aspergillosis." Infection and Immunity 71, no. 5 (May 2003): 2819–26. http://dx.doi.org/10.1128/iai.71.5.2819-2826.2003.

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ABSTRACT Aspergillus fumigatus is the predominant mold pathogen in patients who lack functional innate immunity. The A. fumigatus rhbA gene was first identified as a transcript that was upregulated when the organism was grown in the presence of mammalian cells. To gain insight into the function of rhbA in the growth and pathogenesis of A. fumigatus, we constructed a strain that lacks a functional rhbA gene. The ΔrhbA mutant showed a significant reduction in virulence compared to the virulence of the wild type in a mouse model of invasive aspergillosis. Complementation of the deletion with the wild-type gene restored full virulence. Although the ΔrhbA mutant grew as well as the wild type on solid medium containing the rich nitrogen source ammonium, the growth of the mutant was impaired on medium containing poor nitrogen sources. Like the Saccharomyces cerevisiae rhb1 mutant, the ΔrhbA mutant exhibited increased uptake of arginine. In addition, the ΔrhbA strain underwent asexual development in submerged cultures, even under ammonium-excess conditions. Growth of the mutant with poor nitrogen sources eliminated both the arginine uptake and submerged asexual development phenotypes. The mutant showed enhanced sensitivity to the TOR kinase inhibitor rapamycin. These findings establish the importance of rhbA for A. fumigatus virulence and suggest a role for rhbA in nutrient sensing.

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Zidi-Yahiaoui, Nedjma, Isabelle Mouro-Chanteloup, Anne-Marie D'Ambrosio, Claude Lopez, Pierre Gane, Caroline Le Van Kim, Jean-Pierre Cartron, Yves Colin, and Pierre Ripoche. "Human Rhesus B and Rhesus C glycoproteins: properties of facilitated ammonium transport in recombinant kidney cells." Biochemical Journal 391, no. 1 (September 26, 2005): 33–40. http://dx.doi.org/10.1042/bj20050657.

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The mammalian Rh (Rhesus) protein family belongs to the Amt/Mep (ammonia transporter/methylammonium permease)/Rh superfamily of ammonium transporters. Whereas RhCE, RhD and RhAG are erythroid specific, RhBG and RhCG are expressed in key organs associated with ammonium transport and metabolism. We have investigated the ammonium transport function of human RhBG and RhCG by comparing intracellular pH variation in wild-type and transfected HEK-293 (human embryonic kidney) cells and MDCK (Madin–Darby canine kidney) cells in the presence of ammonium (NH4+/NH3) gradients. Stopped-flow spectrofluorimetry analysis, using BCECF [2′,7′-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein] as a pH-sensitive probe, revealed that all cells submitted to inwardly or outwardly directed ammonium gradients exhibited rapid alkalinization or acidification phases respectively, which account for ammonium movements in transfected and native cells. However, as compared with wild-type cells known to have high NH3 lipid permeability, RhBG- and RhCG-expressing cells exhibited ammonium transport characterized by: (i) a five to six times greater kinetic rate-constant; (ii) a weak temperature-dependence; and (iii) reversible inhibition by mercuric chloride (IC50: 52 μM). Similarly, when subjected to a methylammonium gradient, RhBG- and RhCG-expressing cells exhibited kinetic rate constants greater than those of native cells. However, these constants were five times higher for RhBG as compared with RhCG, suggesting a difference in substrate accessibility. These results, indicating that RhBG and RhCG facilitate rapid and low-energy-dependent bi-directional ammonium movement across the plasma membrane, favour the hypothesis that these Rh glycoproteins, together with their erythroid homologue RhAG [Ripoche, Bertrand, Gane, Birkenmeier, Colin and Cartron (2005) Proc. Natl. Acad. Sci. U.S.A. 101, 17222–17227] constitute a family of NH3 channels in mammalian cells.

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Avornyo, Vincent K., Andrew Manu, David A. Laird, and Michael L. Thompson. "Temperature Effects on Properties of Rice Husk Biochar and Calcinated Burkina Phosphate Rock." Agriculture 11, no. 5 (May 10, 2021): 432. http://dx.doi.org/10.3390/agriculture11050432.

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Rice husk biochar (RHB) and phosphate rock (PR) are locally accessible resources that poor farmers in Sub-Saharan Africa (SSA) can utilize to increase rice productivity. However, biochars are variable depending on feedstock, pyrolysis temperature, and duration. Phosphate rocks from SSA are of low solubility. The goal of this study was to determine whether pyrolysis of rice husk (RH), calcination of PR, and the calcination/pyrolysis of a RHB-PR mixture at 300 °C, 500 °C, and 700 °C can increase formic acid (FA)- extractable phosphorous (P). The properties of these RHBs were compared to the properties of RHB produced through a simple farmer-friendly pyrolysis technique termed “Kun-tan”. Properties of calcinated PR were also compared to the raw PR. Quartz formed from amorphous SiO2 during RH pyrolysis and was the dominant mineral phase in the biochars, irrespective of the pyrolysis temperature. Formic acid-extractable P content, pH, and ash content of the biochars increased with increasing pyrolysis temperature. At 700 °C, FA-extractable P content of the RHB was 219% more than the feedstock. Hydroxyapatite and quartz were the dominant minerals in the PR irrespective of calcination temperature, indicating that hydroxyapatite and quartz were stable to at least 700 °C. Rather, calcination decreased the FA-extractable P content of the PR.

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Han, Ki-Hwan, Hyun-Wook Lee, Mary E. Handlogten, Florence Whitehill, Gunars Osis, Byron P. Croker, William L. Clapp, Jill W. Verlander, and I. David Weiner. "Expression of the ammonia transporter family member, Rh B Glycoprotein, in the human kidney." American Journal of Physiology-Renal Physiology 304, no. 7 (April 1, 2013): F972—F981. http://dx.doi.org/10.1152/ajprenal.00550.2012.

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The ammonia transporter family member, Rh B Glycoprotein (RhBG/Rhbg), is essential for ammonia transport by the rodent kidney, but in the human kidney mRNA but not protein expression has been reported. Because ammonia transport is fundamental for acid-base homeostasis, the current study addressed RhBG expression in the human kidney. Two distinct RhBG mRNA sequences have been reported, with different numbers of consecutive cytosines at nt1265 and thus encoding different carboxy-tails. Sequencing the region of difference in both human kidney and liver mRNA showed eight sequential cytosines, not seven as in some reports. Knowing the correct mRNA sequence for RhBG, we then assessed RhBG protein expression using antibodies against the correct amino acid sequence. Immunoblot analysis demonstrated RhBG protein expression in human kidney and immunohistochemistry identified basolateral RhBG in connecting segment (CNT) and the cortical and outer medullary collecting ducts. Colocalization of RhBG with multiple cell-specific markers demonstrated that that CNT cells and collecting duct type A intercalated cells express high levels of RhBG, and type B intercalated cells and principal cells do not express detectable RhBG. Thus, these studies identify the correct mRNA and thus protein sequence for human RhBG and show that the human kidney expresses basolateral RhBG protein in CNT, type A intercalated cells, and non-A, non-B cells. We conclude that RhBG can mediate an important role in human renal ammonia transport.

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Bishop, Jesse M., Jill W. Verlander, Hyun-Wook Lee, Raoul D. Nelson, Arthur J. Weiner, Mary E. Handlogten, and I. David Weiner. "Role of the Rhesus glycoprotein, Rh B glycoprotein, in renal ammonia excretion." American Journal of Physiology-Renal Physiology 299, no. 5 (November 2010): F1065—F1077. http://dx.doi.org/10.1152/ajprenal.00277.2010.

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Rh B glycoprotein (Rhbg) is a member of the Rh glycoprotein family of ammonia transporters. In the current study, we examine Rhbg's role in basal and acidosis-stimulated acid-base homeostasis. Metabolic acidosis induced by HCl administration increased Rhbg expression in both the cortex and outer medulla. To test the functional significance of increased Rhbg expression, we used a Cre-loxP approach to generate mice with intercalated cell-specific Rhbg knockout (IC-Rhbg-KO). On normal diet, intercalated cell-specific Rhbg deletion did not alter urine ammonia excretion, pH, or titratable acid excretion significantly, but it did decrease glutamine synthetase expression in the outer medulla significantly. After metabolic acidosis was induced, urinary ammonia excretion was significantly less in IC-Rhbg-KO than in control (C) mice on days 2–4 of acid loading, but not on day 5. Urine pH and titratable acid excretion and dietary acid intake did not differ significantly between acid-loaded IC-Rhcg-KO and C mice. In IC-Rhbg-KO mice, acid loading increased connecting segment (CNT) cell and outer medullary collecting duct principal cell Rhbg expression. In both C and IC-Rhbg-KO mice, acid loading decreased glutamine synthetase in both the cortex and outer medulla; the decrease on day 3 was similar in IC-Rhbg-KO and C mice, but on day 5 it was significantly greater in IC-Rhbg-KO than in C mice. We conclude 1) intercalated cell Rhbg contributes to acidosis-stimulated renal ammonia excretion, 2) Rhbg in CNT and principal cells may contribute to renal ammonia excretion, and 3) decreased glutamine synthetase expression may enable normal rates of ammonia excretion under both basal conditions and on day 5 of acid loading in IC-Rhbg-KO mice.

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Handlogten, Mary E., Seong-Pyo Hong, Li Zhang, Allen W. Vander, Marshall L. Steinbaum, Martha Campbell-Thompson, and I. David Weiner. "Expression of the ammonia transporter proteins Rh B glycoprotein and Rh C glycoprotein in the intestinal tract." American Journal of Physiology-Gastrointestinal and Liver Physiology 288, no. 5 (May 2005): G1036—G1047. http://dx.doi.org/10.1152/ajpgi.00418.2004.

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Ammonia metabolism is important in multiple aspects of gastrointestinal physiology, but the mechanisms of ammonia transport in the gastrointestinal tract remain incompletely defined. The present study examines expression of the ammonia transporter family members Rh B glycoprotein (RhBG) and Rh C glycoprotein (RhCG) in the mouse gastrointestinal tract. Real-time RT-PCR amplification and immunoblot analysis identified mRNA and protein for both RhBG and RhCG were expressed in stomach, duodenum, jejunum, ileum, and colon. Immunohistochemistry showed organ and cell-specific expression of both RhBG and RhCG. In the stomach, both RhBG and RhCG were expressed in the fundus and forestomach, but not in the antrum. In the forestomach, RhBG was expressed by all nucleated squamous epithelial cells, whereas RhCG was expressed only in the stratum germinativum. In the fundus, RhBG and RhCG immunoreactivity was present in zymogenic cells but not in parietal or mucous cells. Furthermore, zymogenic cell RhBG and RhCG expression was polarized, with apical RhCG and basolateral RhBG immunoreactivity. In the duodenum, jejunum, ileum, and colon, RhBG and RhCG immunoreactivity was present in villous, but not in mucous or crypt cells. Similar to the fundic zymogenic cell, RhBG and RhCG expression in villous epithelial cells was polarized when apical RhCG and basolateral RhBG immunoreactivity was present. Thus the ammonia transporting proteins RhBG and RhCG exhibit cell-specific, axially heterogeneous, and polarized expression in the intestinal tract suggesting they function cooperatively to mediate gastrointestinal tract ammonia transport.

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Mach, Kathleen E., Kyle A. Furge, and Charles F. Albright. "Loss of Rhb1, a Rheb-Related GTPase in Fission Yeast, Causes Growth Arrest With a Terminal Phenotype Similar to That Caused by Nitrogen Starvation." Genetics 155, no. 2 (June 1, 2000): 611–22. http://dx.doi.org/10.1093/genetics/155.2.611.

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Abstract The Rheb GTPase is most similar in primary sequence to the Ras, Rap, R-Ras, and Ral GTPases, which regulate cell growth and differentiation in many cell types. A likely fission yeast homologue of mammalian Rheb, which we designated Rhb1, was identified by genome sequencing. Our investigation of rhb1 showed that rhb1− cells arrested cell growth and division with a terminal phenotype similar to that of nitrogenstarved cells. In particular, cells depleted of Rhb1 arrested as small, round cells with 1N DNA content, arrested more quickly in low-nitrogen medium, and induced expression of fnx1 and mei2 mRNA, two mRNAs that were normally induced by nitrogen starvation. Since mammalian Rheb binds and may regulate Raf-1, a Ras effector, we tested for functional overlap between Ras1 and Rhb1 in fission yeast. This analysis showed that Ras1 overexpression did not suppress rhb1− mutant phenotypes, Rhb1 overexpression did not suppress ras1− mutant phenotypes, and ras1− rhb1− double mutants had phenotypes equal to the sum of the corresponding single-mutant phenotypes. Hence, there is no evidence for overlapping functions between Ras1 and Rhb1. On the basis of this study, we hypothesize that Rhb1 negatively regulates entry into stationary phase when extracellular nitrogen levels are adequate for growth. If this hypothesis is correct, then Rhb1 and Ras1 regulate alternative responses to limiting nutrients.

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Gómez Gutiérrez, Susana. "Russell y Hegel, una discusión sobre deícticos." Universitas Philosophica 33, no. 66 (March 30, 2016): 113–27. http://dx.doi.org/10.11144/javeriana.uph33-66.rhdd.

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En este trabajo examino las maneras contrapuestas como Bertrand Russell, en varios de sus escritos, y Hegel, en la Fenomenología del espíritu, entienden los llamados deícticos. Intento mostrar que Russell, aun conociendo bien la obra de Hegel, no responde a la propuesta de este último, sino que adopta una posición que lo mantiene atrapado en lo que, desde un punto de vista hegeliano, podría denominarse el autoengaño de la certeza sensible. Al final, sugiero que las diferencias entre Hegel y Russell, a propósito de los deícticos, pueden deberse al

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46

Koizumi, Kazuhisa, Kazunori Takano, Akiko Kaneyasu, Haruko Watanabe-Takano, Emi Tokuda, Tomoyuki Abe, Naoki Watanabe, Tadaomi Takenawa, and Takeshi Endo. "RhoD activated by fibroblast growth factor induces cytoneme-like cellular protrusions through mDia3C." Molecular Biology of the Cell 23, no. 23 (December 2012): 4647–61. http://dx.doi.org/10.1091/mbc.e12-04-0315.

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The small GTPase RhoD regulates actin cytoskeleton to collapse actin stress fibers and focal adhesions, resulting in suppression of cell migration and cytokinesis. It also induces alignment of early endosomes along actin filaments and reduces their motility. We show here that a constitutively activated RhoD generated two types of actin-containing thin peripheral cellular protrusions distinct from Cdc42-induced filopodia. One was longer, almost straight, immotile, and sensitive to fixation, whereas the other was shorter, undulating, motile, and resistant to fixation. Moreover, cells expressing wild-type RhoD extended protrusions toward fibroblast growth factor (FGF) 2/4/8–coated beads. Stimulation of wild-type RhoD-expressing cells with these FGFs also caused formation of cellular protrusions. Nodules moved through the RhoD-induced longer protrusions, mainly toward the cell body. Exogenously expressed FGF receptor was associated with these moving nodules containing endosome-like vesicles. These results suggest that the protrusions are responsible for intercellular communication mediated by FGF and its receptor. Accordingly, the protrusions are morphologically and functionally equivalent to cytonemes. RhoD was activated by FGF2/4/8. Knockdown of RhoD interfered with FGF-induced protrusion formation. Activated RhoD specifically bound to mDia3C and facilitated actin polymerization together with mDia3C. mDia3C was localized to the tips or stems of the protrusions. In addition, constitutively activated mDia3C formed protrusions without RhoD or FGF stimulation. Knockdown of mDia3 obstructed RhoD-induced protrusion formation. These results imply that RhoD activated by FGF signaling forms cytoneme-like protrusions through activation of mDia3C, which induces actin filament formation.

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Sullivan, Harold Clifford, Connie Maridith Arthur, Seema Patel, Jeanne E. Hendrickson, Alan H. Lazarus, and Sean R. Stowell. "Anti-RhD Mediates Loss of RhD Antigen Following Anti-RhD Infusion." Blood 126, no. 23 (December 3, 2015): 3570. http://dx.doi.org/10.1182/blood.v126.23.3570.3570.

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Abstract Background: Previous studies suggest that antibody engagement of red blood cells (RBCs) can result in the actual loss of detectable target antigen in the setting of autoimmune hemolytic anemia. However, many of these situations appear to represent complement mediated-masking of the target antigen. Recent studies suggest that complement-independent antigen-loss can occur in a variety of murine models. However, whether a similar form of RBC antigen loss can occur independent of complement in humans remains unknown. As previous studies suggest that anti-RhD fails to induce significant complement activation following engagement of RhD-positive RBCs, we evaluated the potential impact of anti-RhD antibody injection on RhD antigen levels in a RhD-positive patient being treated for idiopathic thrombocytopenic purpura. Methods: Pre- and post-anti-RhD infusion samples were obtained from the hospital blood bank. Direct antiglobulin tests (DATs) were performed and assessed via flow cytometry on pre- and post-infusion samples. RBCs from pre- and post-infusion were also treated with Warm Autoantibody Removal Medium (WARM; ImmucorGamma, Norcross, GA), a sulphydryl/enzyme reagent based on the ZZAP method, in order to remove bound antibody. Post-WARM treated cells from pre- and post-anti-RhD infusion samples were then used to perform DATs. The presence of RhD antigen was assessed in pre- and post-infusion samples with in vitro anti-RhD, from the same lot number used to treat the patient, followed by anti-IgG and anti-complement as the secondary antibody. To account for the potential impact of bound antibody on antigen detection, antigen levels were also assessed after anti-RhD removal with the WARM method. Finally, in order to determine whether antigen loss was RhD specific, cellano (k) antigen detection was tested using anti-k antibody both pre- and post-WARM treatment. Results: As predicted, the pre-anti-RhD infusion DAT was negative, while the post-anti-RhD injection DAT was positive (MFI 25). Post-WARM treatment, pre-and post-RhD infusion DATs showed minimal reactivity (MFI 3 and 5, respectively with background MFI of 2.7). Reduced RhD antigen levels were observed in post-anti-RhD infusion samples when compared to pre-infusion samples, while no complement could be detected in either pre- or post-anti-RhD infusion samples. The detection of antigen loss post-anti-RhD infusion was even more pronounced after RBCs were treated with WARM to remove previously bound anti-RhD antibody administered in vivo. In contrast, no difference in k antigen level could be detected pre- or post-anti-RhD infusion. As expected, post-WARM treatment, k antigen was no longer detectable pre- or post-anti-RhD infusion samples. Conclusion: These results provide an example of antigen loss in the setting of anti-RhD administration. Moreover, the anti-RhD effect on the RhD-antigen appears to be antigen specific, as the RhD immune globulin did not modulate the k antigen on the same cell. Taken together, these results suggest that anti-RhD can induce the loss of detectable antigen independent of complement and may therefore influence the rate and magnitude of RBC clearance in settings of anti-RhD infusion, incompatible RBC transfusion and autoimmune hemolytic anemia. Disclosures No relevant conflicts of interest to declare.

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Bishop, Jesse M., Hyun-Wook Lee, Mary E. Handlogten, Ki-Hwan Han, Jill W. Verlander, and I. David Weiner. "Intercalated cell-specific Rh B glycoprotein deletion diminishes renal ammonia excretion response to hypokalemia." American Journal of Physiology-Renal Physiology 304, no. 4 (February 15, 2013): F422—F431. http://dx.doi.org/10.1152/ajprenal.00301.2012.

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The ammonia transporter family member, Rh B Glycoprotein (Rhbg), is an ammonia-specific transporter heavily expressed in the kidney and is necessary for the normal increase in ammonia excretion in response to metabolic acidosis. Hypokalemia is a common clinical condition in which there is increased renal ammonia excretion despite the absence of metabolic acidosis. The purpose of this study was to examine Rhbg's role in this response through the use of mice with intercalated cell-specific Rhbg deletion (IC-Rhbg-KO). Hypokalemia induced by feeding a K+-free diet increased urinary ammonia excretion significantly. In mice with intact Rhbg expression, hypokalemia increased Rhbg protein expression in intercalated cells in the cortical collecting duct (CCD) and in the outer medullary collecting duct (OMCD). Deletion of Rhbg from intercalated cells inhibited hypokalemia-induced changes in urinary total ammonia excretion significantly and completely prevented hypokalemia-induced increases in urinary ammonia concentration, but did not alter urinary pH. We conclude that hypokalemia increases Rhbg expression in intercalated cells in the cortex and outer medulla and that intercalated cell Rhbg expression is necessary for the normal increase in renal ammonia excretion in response to hypokalemia.

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49

Han, Ki-Hwan, Kavya Mekala, Venetia Babida, Hye-Young Kim, Mary E. Handlogten, Jill W. Verlander, and I. David Weiner. "Expression of the gas-transporting proteins, Rh B glycoprotein and Rh C glycoprotein, in the murine lung." American Journal of Physiology-Lung Cellular and Molecular Physiology 297, no. 1 (July 2009): L153—L163. http://dx.doi.org/10.1152/ajplung.90524.2008.

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A family of gas-transporting proteins, the Mep/Amt/Rh glycoprotein family, has been identified recently. These are integral membrane proteins, are widely expressed in sites of gas transport, and are known to transport the gaseous molecule, NH3, and recent evidence indicates they can transport CO2. Because the mammalian lung is a critical site for gas transport, the current studies examine the expression of the nonerythroid members of this extended family, Rh B glycoprotein (Rhbg) and Rh C glycoprotein (Rhcg), in the normal mouse lung. Real-time RT-PCR and immunoblot analysis demonstrated both Rhbg and Rhcg mRNA and protein expression, respectively. Immunohistochemistry demonstrated both Rhbg and Rhcg were expressed in bronchial and bronchiolar epithelial cells. Rhbg was expressed by Clara cells, specifically, whereas all bronchial/bronchiolar epithelial cells, with the exception of goblet cells, expressed Rhcg. Rhbg expression was basolateral, whereas Rhcg exhibited apical and intracellular immunolabel, polarized expression similar to that observed in Rhbg- and Rhcg-expressing epithelial cells in other organs. There was no detectable expression of either Rhbg or Rhcg in alveolar endothelial or epithelial cells, in pneumocytes or in vascular tissue. In vitro studies using cultured bronchial epithelial cells confirm Rhbg and Rhcg expression, demonstrate that saturable, not diffusive, transport is the primary mechanism of ammonia/methylammonia transport, and show that the saturable transport mechanism has kinetics similar to those demonstrated previously for Rhbg and Rhcg. These findings suggest Rhbg and Rhcg may contribute to bronchial epithelial cell ammonia metabolism and suggest that they do not contribute to pulmonary CO2 transport.

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Verlander, Jill W., R. Tyler Miller, Amy E. Frank, Ines E. Royaux, Young-Hee Kim, and I. David Weiner. "Localization of the ammonium transporter proteins RhBG and RhCG in mouse kidney." American Journal of Physiology-Renal Physiology 284, no. 2 (February 1, 2003): F323—F337. http://dx.doi.org/10.1152/ajprenal.00050.2002.

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Ammonia is both produced and transported by renal epithelial cells, and it regulates renal ion transport. Recent studies have identified a family of putative ammonium transporters; mRNA for two members of this family, Rh B-glycoprotein (RhBG) and Rh C-glycoprotein (RhCG), is expressed in the kidney. The purpose of this study was to determine the cellular location of RhBG and RhCG protein in the mouse kidney. We generated RhBG- and RhCG-specific anti-peptide antibodies. Immunoblot analysis confirmed that both proteins were expressed in the mouse kidney. RhBG localization with immunohistochemistry revealed discrete basolateral labeling in the connecting segment (CNT) and in the majority of initial collecting tubule (ICT) and cortical collecting duct (CCD) cells. In the outer medullary collecting duct (OMCD) and inner medullary collecting duct (IMCD) only a subpopulation of cells exhibited basolateral immunoreactivity. Colocalization of RhBG with carbonic anhydrase II, the thiazide-sensitive transporter, and the anion exchangers AE1 and pendrin demonstrated RhBG immunoreactivity in all CNT cells and all CCD and ICT principal cells. In the ICT and CCD, basolateral RhBG immunoreactivity is also present in A-type intercalated cells but not in pendrin-positive CCD intercalated cells. In the OMCD and IMCD, only intercalated cells exhibit RhBG immunoreactivity. Immunoreactivity for a second putative ammonium transporter, RhCG, was present in the apical region of cells with almost the same distribution as RhBG. However, RhCG immunoreactivity was present in all CCD cells, and it was present in outer stripe OMCD principal cells, in addition to OMCD and IMCD intercalated cells. Thus the majority of RhBG and RhCG protein expression is present in the same epithelial cell types in the CNT and collecting duct but with opposite polarity. These findings suggest that RhBG and RhCG may play important and cell-specific roles in ammonium transport and signaling in these regions of the kidney.

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